ATPase woes

From:

Sue Danielson <sdaniels@post.its.mcw.edu>

Hello histonetters,
We are working on a research project involving fiber-typing of the
cricopharyngeus muscle in adult human subjects.
Consistently we are finding poor differentiation at pH 9.4, with all fibers
staining darkly. We next attempted to fix this by running the alkaline pH at
10.2. This time all fibers are extremely light (again, no differentation).
We routinely stain skeletal muscle samples such as deltoid and quadriceps at
pH's 9.4, 4.6 and 4.3 with no ill effects.
Our acidic ph's for the atpase procedure still work beautifully on these
cricopharyngeus samples.
Can anyone share any advice or perhaps protocols which work nicely at pH 10.2?
thanks in advance,
Susan Danielson, MS
Neuromuscular Lab Coordinator
Dept Neurology, Medical College of Wisconsin
ph: 414.805.3836
fax: 414.454.7905